title: Mechanosensitivity of astrocytes on optimized polyacrylamide gels analyzed by quantitative morphometry.
abstract: Cells are able to detect and respond to mechanical cues from their environment. Previous studies have investigated this mechanosensitivity on various cell types, including neural cells such as astrocytes. In this study, we have carefully optimized polyacrylamide gels, commonly used as compliant growth substrates, considering their homogeneity in surface topography, mechanical properties, and coating density, and identified several potential pitfalls for the purpose of mechanosensitivity studies. The resulting astrocyte response to growth on substrates with shear storage moduli of G' = 100 Pa and G' = 10 kPa was then evaluated as a function of coating density of poly-D-lysine using quantitative morphometric analysis. Astrocytes cultured on stiff substrates showed significantly increased perimeter, area, diameter, elongation, number of extremities and overall complexity if compared to those cultured on compliant substrates. A statistically significant difference in the overall morphological score was confirmed with an artificial intelligence-based shape analysis. The dependence of the cells' morphology on PDL coating density seemed to be weak compared to the effect of the substrate stiffness and was slightly biphasic, with a maximum at 10-100 µg ml(-1) PDL concentration. Our finding suggests that the compliance of the surrounding tissue in vivo may influence astrocyte morphology and behavior.
authors.affiliation: Department of Physics, Cavendish Laboratory, University of Cambridge, Cambridge, UK.
authors.forename: Pouria
authors.initials: P
authors.lastname: Moshayedi
authors.forename: Luciano da F
authors.initials: Lda F
authors.lastname: Costa
authors.forename: Andreas
authors.initials: A
authors.lastname: Christ
authors.forename: Stephanie P
authors.initials: SP
authors.lastname: Lacour
authors.forename: James
authors.initials: J
authors.lastname: Fawcett
authors.forename: Jochen
authors.initials: J
authors.lastname: Guck
authors.forename: Kristian
authors.initials: K
authors.lastname: Franze
mesh_terms: D000180:Acrylic Resins; D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000502:physiology*; D002448:Cell Adhesion / Q000502:physiology*; D002478:Cells, Cultured; D003198:Computer Simulation; D022001:Focal Adhesions / Q000502:physiology*; D005782:Gels; D006801:Humans; D040542:Mechanotransduction, Cellular / Q000502:physiology*; D008954:Models, Biological; D051381:Rats; D033081:Shear Strength / Q000502:physiology*; D013314:Stress, Mechanical; D000099166:Polyacrylamides
chemical_list: D000180:Acrylic Resins; D005782:Gels; D000099166:Polyacrylamides